膜基质免疫分析法同时检测玉米中伏马菌素B1和呕吐毒素

建立了基于聚偏氟乙烯膜(Polyvinylidene fluoride,PVDF)基质的直接竞争免疫分析法,同时检测玉米中的伏马菌素B1(Fumonisin B1,FB1)及呕吐毒素(Deoxynivalenol,DON)。PVDF膜用甲醇浸湿、激活,用移液器将FB1及DON全抗原点阵于相应的膜反应区,同时采用三聚氰氯法和高碘酸钠法分别制备抗FB1、抗DON的辣根过氧化酶(Horseradish peroxidase,HRP)标抗体(monoclonal antibody,McAb),以直接竞争免疫检测方法的模式实现同时检测玉米中的FB1及DON。该方法对于FB1和DON的检出限分别为2.5和50μg/L,样品前处理简单,检测时间15 min,可肉眼辨别结果,随机检测了30份市售玉米样品,并与市售ELISA试剂盒进行方法学比较,结果无明显差异。

Simultaneous Detection of Fumonisin B1 and Deoxynivalenol in Corn Samples Using a Polyvinylidene Fluoride Membrane-based Immunoassay

A Polyvinylidene fluoride(PVDF) membrane-based direct competition immunoassay for simultaneous detection of fumonisin B1(FB1)and deoxynivalenol(DON) in corn samples was developed.PVDF membrane was soaked and activated in methanol,and then the conjugates(FB1-ovalbumin(OVA) and DON-cBSA) were spotted on the membrane reaction zone using the tip of dispenser.Horseradish peroxidase(HRP) was successfully coupled to anti-FB1 monoclonal antibody(McAb) and anti-DON McAb by cyanuric chloride method and periodate method respectively.Simultaneous detection of fumonisin B1 and deoxynivalenol in one membrane in the system was carried out under direct competition immunoassay mode.The detection limits for fumonisins B1 and deoxynivalenol were 2.5 and 50 μg/L,respectively.With very simple sample pre-treatment,a batch of extracted samples was analyzed within 15 min.The concentration and type of mycotoxins in samples were determined visibly by the naked eye.Fumonisins B1 and deoxynivalenol in 30 corn samples were screened by the PVDF membrane-based immunoassay,the results showed no significant difference with the ELISA kit.